rp2 prep stained squash of cell from plant root tip Flashcards

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1
Q

how to prep squashes of cells from plant root tip

A

cut thin slice of root tip using scalpel
mount onto slide
soak root tip in hcl rinse
stain for dna
lower coverslip using mounted needle at 45 without trapping air bubbles
squash by firmly pressing down on glass slip but dont push sideways

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2
Q

why are root tips used

A

where dividing cells found and where mitosis occurs

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3
Q

why is stain used

A

to distinguish chromosomes

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4
Q

why press down on coverslip

A

spread cells = single layer of cells
so light passes through to make chromosomes visible

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5
Q

why not push coverslip sideways

A

avoids rolling cells together and breaking chromosomes

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6
Q

why soak roots in acid

A

separates cells
allows stain to diffuse into cell
allow cells to be squashed more easily
stops mitosis

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7
Q

how to set up and use microscope

A

clip slide onto stage and turn on light
select lowest power objective lense
use coarse focusing dial to move stage close to lens
turn coarse focusing dial to move stage away from lens till image comes into focus
just fine focusing dial to get clear image
swap to higher power objective lens and refocus

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8
Q

appearance of prophase

A

chrom visible bc condensing
randomly arranged as not attached to SF

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9
Q

metaphase appearance

A

chrome line up at equator
bc attaching to spindle

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10
Q

anaphase appearance

A

chrom at poles of spindle
chrom v shaped bc being pulled apart at their centromeres by SF

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11
Q

telophase

A

chrom in two sets
one at each pole

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12
Q

mitotic index

A

prop of cells undergoing mitosis divide by total cells in sample

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13
Q

how to determine reliable MI

A

count cells in mitosis
count only whole cells
divided by total no of cells
repeat w 5 fields of view selected randomly
calc mean

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